6XFM
Molecular structure of the core of amyloid-like fibrils formed by residues 111-214 of FUS
ELECTRON MICROSCOPY
| Sample |
|---|
| FUS low complexity sequence |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | HELICAL ARRAY |
| Vitrification Instrument | LEICA PLUNGER |
| Cryogen Name | ETHANE |
| Sample Vitrification Details | Preblot for 10 seconds and blot for 5 seconds before plunging |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | SINGLE PARTICLE |
| Number of Particles | 275520 |
| Reported Resolution (Å) | 2.62 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | 3D refinement and post-processing were performed with 21 (screw) symmetry |
| Refinement Type | |
| Symmetry Type | POINT |
| Point Symmetry | C1 |
| Axial Symmetry | C2 |
| Axial Rise | 4.8 |
| Angular Rotation | -2 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 | ||||
| Refinement Space | |||||
| Refinement Protocol | OTHER | ||||
| Refinement Target | |||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | Manually generated model was fit into the density using PHENIX and UCSF Chimera. Further refinements were performed using Xplor-NIH. | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K2 SUMMIT (4k x 4k) | ||||||||
| Electron Dose (electrons/Å**2) | 47 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | |
| Temperature (Kelvin) | |
| Microscope Model | FEI TITAN KRIOS |
| Minimum Defocus (nm) | 1000 |
| Maximum Defocus (nm) | 2500 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | |
| Nominal Magnification | 130000 |
| Calibrated Magnification | |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 300 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| PARTICLE SELECTION | RELION | 3.0 |
| IMAGE ACQUISITION | SerialEM | |
| CTF CORRECTION | RELION | 3.0 |
| MODEL FITTING | Coot | |
| INITIAL EULER ASSIGNMENT | RELION | 3.0 |
| FINAL EULER ASSIGNMENT | RELION | 3.0 |
| CLASSIFICATION | RELION | 3.0 |
| RECONSTRUCTION | RELION | 3.0 |
| MODEL REFINEMENT | PHENIX | |
| MODEL REFINEMENT | UCSF Chimera | |
| MODEL REFINEMENT | X-PLOR | Xplor-NIH |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| NONE | Gctf | 499206 | 499206 of particles were extracted from the 58185 fibril segments using a 400-pixel box size and 91.6% overlap. | |
